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Assay Method Information

Assay Name:  Fluorimetric Assay
Description:  Inhibitory constants (Ki) and IC50 values were determined under saturating substrate conditions by using a fluorimetric assay (components were purchased separately). This assay system allows detection of a fluorescent signal upon SIRT6 deacetylation of the fluorescent substrate peptide (AMC-p53) with amino acids 379-382 (Arg-His-Lys-LysAc). Sirtuin deacetylation of the substrate renders the fluorophore-substrate bond susceptible to treatment with lysyl endopeptidase. Trypsin cleavage releases the fluorophore, resulting in an increase in fluorescence. Fluorescence intensity was measured on a Synergy HT Multi-Mode microplate fluorescence reader.
Affinity data for this assay
 

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Last update November 1, 2007
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